Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Enzyme-linked immunosorbent assay kit for detecting ivermectin and abamectin and application of enzyme-linked immunosorbent assay kit

An enzyme-linked immunosorbent reagent, ivermectin technology, which is applied to measurement devices, analyzes materials by chemical reaction, and analyzes materials by observing the impact on chemical indicators. The problems of rapid sample detection, complex processing process, and long time-consuming can achieve the effect of convenient and easy inspection methods, low pretreatment requirements, and convenient use.

Active Publication Date: 2019-03-29
BEIJING KWINBON BIOTECH
View PDF9 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods can simultaneously perform multi-residue qualitative and quantitative analysis of abamectins, but require expensive instruments and specialized technicians. The sample pretreatment process is complex, costly, and time-consuming, and it is difficult to meet the rapid detection of a large number of samples and on-site samples. needs

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Enzyme-linked immunosorbent assay kit for detecting ivermectin and abamectin and application of enzyme-linked immunosorbent assay kit
  • Enzyme-linked immunosorbent assay kit for detecting ivermectin and abamectin and application of enzyme-linked immunosorbent assay kit
  • Enzyme-linked immunosorbent assay kit for detecting ivermectin and abamectin and application of enzyme-linked immunosorbent assay kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] The preparation of embodiment 1 kit components

[0027] 1. Synthesis of ivermectin hapten (synthetic route see appendix figure 1 )

[0028] Take 1.0g of ivermectin, add 50mL of dichloromethane to dissolve, add 0.45g of pyridinium dichromate, add 0.5mL of glacial acetic acid, stir at 60°C for 4h, stop the reaction, evaporate to dryness, add water 100mL, add ethyl acetate 150mL× 3. Extracted three times, combined the organic phases, concentrated and evaporated to dryness, put on a silica gel column, and eluted with ethyl acetate / petroleum ether (v / v, 1 / 5) to obtain 0.9 g of keto-ivermectin with a yield of 90.2%;

[0029] Take 0.9g of keto-ivermectin, add 60mL of tetrahydrofuran to dissolve, add 0.24g of 1,3-dioxane-2-ethylmagnesium bromide, stir at room temperature for 3h, stop the reaction, add 60mL of ice water, add ethyl acetate Ester 100mL×3, extracted three times, combined the organic phases, evaporated to dryness, and recrystallized from ethyl acetate / n-hexane (v / ...

Embodiment 2

[0062] Embodiment 2 detects the formation of the ELISA kit of ivermectin and abamectin

[0063] An ELISA kit for detecting ivermectin and avermectin was set up to include the following components:

[0064] (1) A microtiter plate coated with an ivermectin-conjugated antigen;

[0065] (2) 6 bottles of ivermectin standard solution, the concentrations were 0 μg / L, 1 μg / L, 3 μg / L, 9 μg / L, 27 μg / L, 81 μg / L;

[0066] (3) Ivermectin monoclonal antibody working solution;

[0067] (4) goat anti-mouse anti-antibody labeled with horseradish peroxidase;

[0068] (5) Substrate chromogenic solution is made up of A liquid and B liquid, and A liquid is carbamide peroxide, and B liquid is tetramethylbenzidine;

[0069] (6) The stop solution is 2mol / L sulfuric acid;

[0070] (7) The washing solution has a pH value of 7.4 and contains 0.5% to 1.0% Tween-20, 0.01‰ to 0.03‰ sodium azide, and 0.1 to 0.3mol / L phosphate buffer;

[0071] (8) The complex solution is a phosphate buffer solution with...

Embodiment 3

[0072] The detection of ivermectin in the sample of embodiment 3

[0073] 1. Sample pretreatment

[0074] (1) Animal tissue

[0075] Homogenize fresh tissue samples with a homogenizer; weigh 4.0g±0.05g of homogenized tissue samples into a 50mL polystyrene centrifuge tube, add 4mL of methanol, shake with an oscillator for 5min, and mix well; 3000g room temperature (20- Centrifuge at 25°C / 68-77°F) for 5min; Measure 2mL of supernatant into a 10mL polystyrene centrifuge tube, add 3mL of acetonitrile and 1mL of n-hexane, then add 1g of neutral alumina, vortex for 1min with a vortexer, Mix well; centrifuge at 3000g at room temperature (20-25°C / 68-77°F) for 5min; remove the upper layer of n-hexane, take 1mL of the clear solution into a 10mL clean and dry glass test tube, and place it in a water bath with nitrogen at 50-60°C (122-140°F) Blow dry under flow; add 100 μL of methanol, vortex for 30 seconds with a vortex, then add 900 μL complex solution, vortex for 30 seconds with a vor...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention provides an enzyme-linked immunosorbent assay kit for detecting ivermectin and abamectin. The enzyme-linked immunosorbent assay kit comprises an ELISA plate coated with an ivermectin coupling antigen, an ivermectin monoclonal antibody, an enzyme-labeled antibody, an ivermectin standard substance solution, a solution substrate colored solution, a stopping solution, a washing solutionand a combination solution. The invention also discloses a method for detecting the residual quantity of ivermectin and abamectin by using the enzyme-linked immunosorbent assay kit. The method comprises the following steps: firstly performing pretreatment on a sample, then detecting the sample by using a kit, and finally analyzing the detection result. The enzyme-linked immunosorbent assay kit provided by the invention can be used for detecting the residual quantities of ivermectin and abamectin in animal tissues, milk and water. The enzyme-linked immunosorbent assay kit provided by the invention is simple and convenient in operation, low in cost and high in sensitivity, which can be used for the field monitoring and the screening of a large number of samples.

Description

technical field [0001] The present invention relates to ELISA detection technology, in particular to an ELISA kit for detecting ivermectin and avermectin, which is especially suitable for ivermectin and avermectin in animal tissues, milk and water quality Detection of mycocin residues. Background technique [0002] Both ivermectin and avermectin belong to the avermectin class of drugs. As a kind of antiparasitic drugs, avermectins have a novel chemical structure, unique mechanism of action, strong insecticidal activity and wide insecticidal spectrum, which have been hailed as a major breakthrough in the research of antiparasitic drugs in the past 20 years. A new type of macrolide antiparasitic drug with the advantages of broad spectrum, high efficiency, small dosage and safety, etc., has high-efficiency repellent and killing effects on nematodes, insects and mites. Although its dosage is relatively small, the residues in animal-derived foods may cause potential harm to hum...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N21/78
CPCG01N21/78G01N33/577
Inventor 何方洋万宇平王兆芹崔廷婷韩深冯才伟朱亮亮李楠申梁刘二战
Owner BEIJING KWINBON BIOTECH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com